Hepatocellular Vacuolization in Rabbits: Effects of Feed Restriction, Orchiectomy and Ovariectomy
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Previous studies have reported that striking variations in hepatocellular vacuolization occurs in rabbits and the magnitude of vacuolization correlated independently with weight and sex. The current study evaluated the effects of feed restriction and gonadectomy on this hepatocellular vacuolization. For 28 days, rabbits were fed either ad libitum (ad lib group), 100% of the National Research Council's recommended feed intake required for growth (100% group), or 50% of the NRC recommended feed intake required for growth (50% group). Feed consumption, weight gain, final body weight, absolute liver weight, and relative liver weight were not significantly different between the ad lib and 100% groups. Values for these parameters for both groups were significantly greater than for the 50% group. Rabbits in the 50% group had significantly less hepatocellular vacuolization than rabbits in the 100% group. Hepatocellular vacuolization in the 100% group did not differ from rabbits fed ad libitum. Hepatocellular vacuolization in the ad lib group was greater than in the 50% group but this difference was not significant. Ovariectomy and orchiectomy did not significantly alter hepatocellular vacuolization in either female or male rabbits, respectively, that were fed ad libitum for 28 days. However, intact females had significantly greater hepatocellular vacuolization than either intact or orchiectomized males. Conversely, hepatocellular vacuolization in ovariectomized females was not significantly different from that in intact and orchiectomized males. There were no significant differences in feed consumption, weight gain, final body weight, and absolute and relative liver weights among these intact and gonadectomized groups. Results of these studies indicate feed consumption can affect the degree of hepatocellular vacuolization in rabbits.(ABSTRACT TRUNCATED AT 250 WORDS)Keywords:
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In order to clarify growth effects of gamma-ray irradiation on the third-stage larvae of Angiostrongylus cantonensis, snails naturally infected with larvae of A. cantonensis were irradiated with a cobalt-60 source, and the total dosage of exposure was 0.1 kGy. Third-stage larvae were collected from each group of irradiated and non-irradiated snails and then administered to rats. A significant reduction was observed in the number of recovery worms from rats sacrificed at 45 days after infection compared with the control group. Moreover, there was an increase in the percentage of female worms, and the body length of the irradiated group's worms was significantly shorter than that of the control group. In ultrastructural observations by transmission electron microscope, the cuticle was thickened and the subcuticle showed partial vacuolization and separation from muscle in the irradiated group. In addition, the muscles myofibril became shorter, the microvilli of intestinal mucosa were shorter and irregular, the terminal web disappeared, and the phagocytic vesicles, mitochondria, lysosomes and secretion granules decreased in number, respectively. Focusing on the mitochondria of the epithelial cells of the intestine, there were degenerative changes in the irradiated group showing vacuolization. The gonads of female worms recovered from the irradiated group revealed degenerative vacuolization, and the gonads of male worms could not be found.
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Vacuolization has been known for two decades to occur in the cytoplasm and over the nuclei of the erythroid and myeloid precursors in bone marrows of patients with acute alcoholism. Electron microscopic examination of the marrows from four acute alcohol-intoxicated subjects disclosed that the vacuoles are present only in the cytoplasm and free of organized structure. Surface invagination of the cell membrane of erythroblasts leads to endocytosis and consequent vacuole formation. Cytoplasmic vacuolization of bone marrow cells was reproduced in vitro in 8 of 12 bone marrows from normal individuals when incubated for 6 hours or more in nutrient medium containing alcohol. The critical alcohol concentration for vacuolization was 62.5 mg/dl. The proportion of cells developing vacuoles appeared to correlate with the concentration of alcohol particularly above levels of 250 mg/dl.
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// Andrey V. Shubin 1,2,3,* , Ilya V. Demidyuk 1,* , Alexey A. Komissarov 1 , Lola M. Rafieva 1 and Sergey V. Kostrov 1 1 Laboratory of Protein Engineering, Institute of Molecular Genetics, Moscow, Russia 2 Laboratory of Chemical Carcinogenesis, N.N. Blokhin Russian Cancer Research Center, Moscow, Russia 3 Laboratory of Biologically Active Nanostructures, N.F. Gamaleya Institute of Epidemiology and Microbiology, Moscow, Russia * These authors contributed equally to this work Correspondence to: Andrey V. Shubin, email: // Keywords : regulated cell death, vacuolization, microbial toxins, viruses Received : February 27, 2016 Accepted : June 06, 2016 Published : June 17, 2016 Abstract Cytoplasmic vacuolization (also called cytoplasmic vacuolation) is a well-known morphological phenomenon observed in mammalian cells after exposure to bacterial or viral pathogens as well as to various natural and artificial low-molecular-weight compounds. Vacuolization often accompanies cell death; however, its role in cell death processes remains unclear. This can be attributed to studying vacuolization at the level of morphology for many years. At the same time, new data on the molecular mechanisms of the vacuole formation and structure have become available. In addition, numerous examples of the association between vacuolization and previously unknown cell death types have been reported. Here, we review these data to make a deeper insight into the role of cytoplasmic vacuolization in cell death and survival.
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Objectives: To observe the effects of Moringa Oleifera leaves extract on BPA induced changes (Diameter of hepatocytes and vacuolization) in liver of rats.
Study Design: Experimental study, conducted at Post Graduate Medical Institute.
Methodology: This study was performed on 32 adult rats for seven weeks. They were divided into 4 equal groups A, B, C and D. Group A was control received corn oil only. Group B, received BPA only 50mg/kg/bw. Group C and D received BPA 50mg/kg along with MoLE 250mg and 500mg. Liver was removed and was fixed in 10% formalin. To observe the effect of BPA and MoLE, slides were prepared for histological examination. For Hepatocytes, diameter and vacuolization was observed. The statistical analysis of results was done by using SPSS 21.
Result: In group B, vacuolization (87.5% of animals) and statistical significant increase in mean diameter (19.7±1.3) of hepatocytes was seen. Presence of vacuoles has presumably lead to an increase in diameter . However in Groups C 50% of animals showed vacuolization and mean mean hepatocyte diameter was 17.0±1.1. In Group D 25% of animals showed vacuolization and mean mean hepatocyte diameter decreased to14.6±1 after administration of MoLE.
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Previous studies have reported that striking variations in hepatocellular vacuolization occurs in rabbits and the magnitude of vacuolization correlated independently with weight and sex. The current study evaluated the effects of feed restriction and gonadectomy on this hepatocellular vacuolization. For 28 days, rabbits were fed either ad libitum (ad lib group), 100% of the National Research Council's recommended feed intake required for growth (100% group), or 50% of the NRC recommended feed intake required for growth (50% group). Feed consumption, weight gain, final body weight, absolute liver weight, and relative liver weight were not significantly different between the ad lib and 100% groups. Values for these parameters for both groups were significantly greater than for the 50% group. Rabbits in the 50% group had significantly less hepatocellular vacuolization than rabbits in the 100% group. Hepatocellular vacuolization in the 100% group did not differ from rabbits fed ad libitum. Hepatocellular vacuolization in the ad lib group was greater than in the 50% group but this difference was not significant. Ovariectomy and orchiectomy did not significantly alter hepatocellular vacuolization in either female or male rabbits, respectively, that were fed ad libitum for 28 days. However, intact females had significantly greater hepatocellular vacuolization than either intact or orchiectomized males. Conversely, hepatocellular vacuolization in ovariectomized females was not significantly different from that in intact and orchiectomized males. There were no significant differences in feed consumption, weight gain, final body weight, and absolute and relative liver weights among these intact and gonadectomized groups. Results of these studies indicate feed consumption can affect the degree of hepatocellular vacuolization in rabbits.(ABSTRACT TRUNCATED AT 250 WORDS)
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Marked variation in hepatocellular vacuolization was present in New Zealand white rabbits used as controls in 28-day and 91-day percutaneous studies conducted at 5 different laboratories. Vacuoles in hepatocytes of alcohol-fixed and formalin-fixed livers contained periodic acid-Schiff (PAS)-positive material which was removed with diastase digestion, indicating the presence of glycogen. The magnitude of hepatocyte vacuolization was subjectively assessed by light microscopy using 5 histologic grades. Quantitative measurements of hepatocyte perimeter and lobule radius for representative liver sections of each histologic grade corroborated the different grades used. Factors associated significantly with the degree of vacuolization were sex (females were affected more severely than males), body weight, relative liver weight, and the laboratory conducting the study. Also apparent were variations in mean severity of hepatocyte vacuolization between studies conducted at the same laboratory, and variation in severity of vacuolization within individual studies. Duration of the study and season had no significant association with the degree of vacuolization. Marked variation of hepatocellular vacuolization due to glycogen accumulation must be recognized when evaluating results of toxicity testing in rabbits.
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Tissue culture cells were exposed to supernatants of Helicobacter pylori for 24 h at 37 degrees C in the presence of various quantities of urea. In the normal human stomach the concentration of urea is less than or equal to 4 mmol/l, and in the presence of this low concentration up to 10% of Vero cells showed intracellular vacuolization. In the presence of 7.5 mmol/l urea, 25% of the cells showed vacuolization. With 30 mmol/l urea, the final pH was 7.6, indicating that vacuolization was not due to change of pH. The first report of vacuolization of tissue culture cells by H. pylori was in a system without added urea but with concentrated bacterial supernatant; 30% of H. pylori strains demonstrated a cytotoxic effect. In those experiments fetal calf serum was used; it contains 6 mmol/l urea but was used at a concentration of 10%. A urease inhibitor, acetohydroxamic acid, caused a 75% drop in the number of cells showing vacuolization, and ammonia caused vacuolization. Thus the urea of H. pylori probably causes this vacuolization.
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In mucopolysaccharidosis type I (MPS I; alpha-L-iduronidase deficiency), glycosaminoglycans (GAGs) accumulate in different cell types, causing characteristic vacuolization. Hematopoietic cell transplantation (HCT) and enzyme replacement therapy (ERT) both aim to restore tissue morphology by delivering alpha-L-iduronidase to the deficient cells. The authors investigated the efficacy of both therapies on dermal fibroblast morphology in 12 patients by electron microscopy of repeated skin biopsies before and during 2 years of ERT as well as before and 6 months after HCT. Cell vacuolization was rated according to a semi-quantitative scoring system. At baseline all patients showed an increased vacuolization score as compared to controls. In addition the vacuolization score was significantly higher in patients with the severe phenotype of the disease (n = 7) compared to patients with attenuated phenotypes (n = 5) (p = .009). After initiation of ERT a significant decrease in cell vacuolization was observed (p = .012). However, the response rate varied among patients, as the vacuolization score remained high during the first year of ERT in 3 patients with the severe phenotype. In all patients who received a successful HCT (n = 3) only minimal disturbances in cell morphology were observed afterward. In conclusion, both ERT and HCT are capable of restoring, at least partially, dermal fibroblast morphology in MPS I.
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